环磷酰胺对BALB／C（H-2^d）雌性成年小鼠卵巢形态结构与功能的影响

Effects of chemotherapy before hematopoietic stem cell transplantation on the ovarian structure and function

目的探讨造血干细胞移植前环磷酰胺预处理对卵巢的损伤作用。方法将60只BALB/C(H-2d)小鼠随机分为A组(20只),不给予任何处理;B组(20只),经腹腔内注入生理盐水0.5m l;C组(20只),以环磷酰胺溶解于生理盐水后,按380mg/kg剂量一次性腹腔注射。C组分别于预处理后15d、(C1组)、30d(C2组)各取10只小鼠处死,A组(A1组、A2组)和B组(B1组、B2组)分别于第2、4个间情期各取10只小鼠处死,分别在光镜和电镜下观察卵巢组织形态学改变,计数始基卵泡、初级卵泡、窦状卵泡和排卵前卵泡。放免法检测血清E2、P、T、FSH、LH、PRL。结果C组小鼠于预处理后出现精神萎靡、体重下降、阴道上皮细胞失去周期性变化等表现。卵巢体积缩小,光镜下颗粒细胞变性、坏死,发育期卵泡数减少,闭锁卵泡增加(P<0.01),但始基卵泡数正常。电镜下可见卵泡透明带完整,卵母细胞、卵丘细胞和颗粒细胞均受到不同程度损伤。C组E2、P显著低于空白组和对照组,FSH、LH水平则显著高于A组和B组,(P<0.01);T、PRL水平在各组之间无显著性差异,预处理30d小鼠较15d损伤程度减轻。结论造血干细胞移植前化学药物预处理可造成卵巢结构与功能的损伤,但这种损伤未影响始基卵泡,卵巢尚有一定储备功能,随距药物作用时间的延长其结构与功能还可在一定程度上得以恢复。

Objective ： This article was to study the changes of ovarian function and structure of mice after conditioning therapy by used chemotherapeutic agent before hematopoietic stem cell transplantation. Methods： Sixty female BALA/C （H -2d） mice were random divided into group A （n = 20） ： blank group; group B （n = 20） ： control group; and group C （n = 20） ： conditioning regiments u- sing cyclophosphamide; Mice were sacrificed respectively at 15 day and 30 day after conditioning regiments using cyclophosphamide （ Group A and B were sacrificed respectively on the day of the second diestrus and the fourth diestrus）. The ovarian tissues were ob- served by light microscope and electron microscope. Blood samples were obtained and serous E2, P, FSH, LH, T, PRL levels were assayed by radioimmunoassay （RIA）. Results： After conditioning regiments used cyclophosphamide, mice were observed to be inaction, body weight dropping and disappearance of the estrous cyclicity. The ovarian volumes become small. The numbers of growing follicles significantly decreased and the numbers of the atretic follicles significantly increased （ P 〈 0. 01 ）. However, primordial follicles＇ numbers had not any difference in all groups. The necrosis of granulosa cells was observed in ovary by light - microscope. Follicular granulosa cells degeneration and necrosis were observed in follicles by electronic microscope. Apoptosis of cells could be observed in the group C, but the zona pellucida was integrated. The serous levels of E2 and P （ P 〈0.01 ） were lower while serous FSH and LH levels were significantly higher （ P 〈 0. 01 ） in the mice of group C. There was no difference in the T and PRL levels in all subjects. Conclusions： The damage of ovarian function and structure of mice results from conditioning therapy by used chemotherapeutic agent before hematopoietic stem cell transplantation. But the the primordial follicles and the cells in the ovarian interstitium were not damaged indicated that the ovarian damage could recover.